WO2003082027A1 - Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif - Google Patents

Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif Download PDF

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Publication number
WO2003082027A1
WO2003082027A1 PCT/JP2002/003177 JP0203177W WO03082027A1 WO 2003082027 A1 WO2003082027 A1 WO 2003082027A1 JP 0203177 W JP0203177 W JP 0203177W WO 03082027 A1 WO03082027 A1 WO 03082027A1
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Prior art keywords
lactic acid
strain
lactobacillus
bacteria
acid bacteria
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PCT/JP2002/003177
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English (en)
Japanese (ja)
Inventor
Yasuhiro Koga
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Frente International Co., Ltd.
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Application filed by Frente International Co., Ltd. filed Critical Frente International Co., Ltd.
Priority to PCT/JP2002/003177 priority Critical patent/WO2003082027A1/fr
Priority to CNB02828691XA priority patent/CN1309316C/zh
Priority to DK02708726.1T priority patent/DK1498039T3/da
Priority to US10/508,340 priority patent/US7674613B2/en
Priority to KR1020047015110A priority patent/KR100843020B1/ko
Priority to EP02708726A priority patent/EP1498039B1/fr
Priority to JP2003579579A priority patent/JP4203855B2/ja
Priority to AU2002243005A priority patent/AU2002243005A1/en
Priority to TW92107146A priority patent/TW200305370A/zh
Publication of WO2003082027A1 publication Critical patent/WO2003082027A1/fr
Priority to HK05106059A priority patent/HK1073230A1/xx

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
    • A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
    • A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
    • A23G3/364—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
    • A23G3/366—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G4/00—Chewing gum
    • A23G4/06—Chewing gum characterised by the composition containing organic or inorganic compounds
    • A23G4/12—Chewing gum characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
    • A23G4/123—Chewing gum characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/179—Sakei
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the present invention relates to a pharmaceutical product for total oral care (living bacterial preparation) or a food containing lactic acid bacteria for the purpose of preventing or treating gingivitis, periodontitis, periodontal disease, caries and bad breath, comprising lactic acid bacteria as an active ingredient Regarding use.
  • a pharmaceutical product for total oral care living bacterial preparation
  • a food containing lactic acid bacteria for the purpose of preventing or treating gingivitis, periodontitis, periodontal disease, caries and bad breath, comprising lactic acid bacteria as an active ingredient Regarding use.
  • periodontal disease has been pointed out not only as a chronic infection of gingival tissues, but also as a risk of causing cardiovascular infarction and blood vessel rupture due to aneurysms. Furthermore, it is attracting attention as a risk factor for diabetes and premature birth.
  • treatment by dental technology that is, administration of bactericide to the affected area, surgery, use of antibiotics by oral administration, etc. ing.
  • long-term continuous administration of bactericides and antibiotics for treatment creates new resistant bacteria due to their use and has many problems such as the occurrence of side effects caused by drugs. Is currently not established.
  • O3 ⁇ 4s orafo is expressed or produced) transferase) P, a factor that suppresses harmful activity and caries growth, and the above lactic acid bacteria that inhibit hydrogen peroxide production have been investigated.
  • more than 400 types of bacteria live in the human oral cavity and the number of bacteria has reached 10 billion.
  • 10 8 ⁇ :! 0 9 CFU / ml level of bacteria is detected. Therefore, a complex flora (oral bacterial flora) is formed in the human oral cavity, and it is effective for bruises with results obtained in a simple in vitro system for the prevention and treatment of periodontal diseases and caries.
  • an object of the present invention is to provide a lactic acid strain capable of preventing the onset, recurrence, and relapse of periodontal disease caused by periodontal disease bacteria and caries bacteria and Z or caries.
  • Another object of the present invention is to provide a viable bacterial preparation containing the above lactic acid strain and a food containing lactic acid bacteria.
  • the present inventor has searched for various lactic acid bacteria belonging to the genus Lactobacillus from the above viewpoint, and found that Lactobacillus salivarius has the above-mentioned characteristics. completed.
  • the present invention provides a live bacterial preparation or a food containing a lactic acid bacterium containing the lactic acid bacterium Ratatobacillus salinorius as an active ingredient.
  • the live bacteria preparation or lactic acid bacterium-containing food of the present invention is used to normalize oral flora, prevent or treat gingivitis, periodontitis and periodontal disease, prevent or treat caries, prevent bad breath and remove bad breath It can be used for
  • the present invention provides Lactobacillus salivarius TI 2711 strain (FERM BP-7974) as a particularly preferred strain of Lactobacillus salivarius, and microbial cells and dried microbial cells thereof.
  • the viable preparation and lactic acid bacteria-containing food of the present invention described above particularly preferably include Lactobacillus salivarius TI 2711 strain (FERM BP-7974) as Lactobacillus salivarius.
  • the present invention also relates to the use of the lactic acid bacteria Ratatobacillus sarivarius TI 2711 strain (FERM BP-7974) in the production of a live bacterial preparation of Lactobacillus salivarius TI 2711 strain (FERMBP-7974) or a food containing lactic acid bacteria, and the lactic acid bacterium Lactobacillus
  • the present invention provides a composition comprising a Sari barius TI 2711 (FERM BP-7974) strain and an active ingredient having a mechanism of action different from that of the strain.
  • FIG. 1 is a graph showing the inhibitory effect (in vivo) of L. salivarius ⁇ l 2711 strain on the mouse oral cavity JR gingivalis JCM 8525 strain.
  • FIG. 2 is a graph showing the inhibitory effect (in vivS) of the L. salivarius TI 2711 strain on the mouse oral S. mutans MT 8148 strain.
  • Fig. 3 is a graph showing changes in the total number of bacteria in the human oral cavity.
  • Fig. 4 is a graph showing changes in the number of human oral periodontal disease bacteria (BPAE).
  • FIG. 5 is a graph showing changes in the number of Streptococcus mutans within the human space.
  • FIG. 6 is a graph showing changes in the number of human oral lactic acid bacteria.
  • FIG. 7 is a graph showing changes in the pH of human saliva.
  • FIG. 8 is a graph showing changes in the amount of insoluble glucan in human saliva.
  • FIG. 9 is a graph showing the results of measuring the human odor with a halimeter.
  • FIG. 10 is a diagram showing an outline of the protocol of Experiment 1 and Experiment 2 in Test Example 2.
  • FIG. 11 is a diagram showing an outline of the protocol of the clinical trial in Test Example 3. BEST MODE FOR CARRYING OUT THE INVENTION
  • the present inventor tested a large number of stored strains of the genus Lactobacillus (30 strains) isolated from the oral cavity of a healthy person according to the following procedure. The optimal strain was selected.
  • Lactobacillus salivarius ⁇ 2711 selected in ⁇ 4 are as follows.
  • Nitrate reducing ability One
  • Trehalose + Based on the above bacteriological properties, classification of Virgie's Manual ⁇ Ob ⁇ Systematics ⁇ Batterologic 1, Vol. 2, (1986) and the World of Enterobacteria, Mitsuoka Chitose, Chosokusha, (1980) According to the standards, the strains of the present invention selected in the above 1 to 4 were the same as Lactobacillus salivarius, and the strain was named Lactooacilius salivarius TI 2711 strain.
  • the lactic acid strain of the present invention ⁇ cto> a 3 ⁇ 4w sa / ari i7sTI 2711 strain is ingested by a host such as a human to the gum tissue, periodontal pockets and teeth of periodontal disease bacteria and caries bacteria. It can suppress or prevent growth and suppress or eliminate pathogenic bacteria present in the oral cavity. This provides an extremely effective means for preventing or treating periodontal disease and caries, which can be called national illness.
  • Such interactions between the host and oral microorganisms have been widely observed in nature, and are understood as symbiosis, parasitism, or antagonism in the relationship between the host and the microorganism.
  • the lactic acid strain Lactobacillus salivarius ⁇ 2 W) of the present invention can be administered as an active ingredient as it is or as a single agent together with an appropriate additive, and also with other active ingredients such as the strain.
  • Different oral care drugs of different It can be administered at the same time or as a mixture.
  • As its administration form for example, powders, granules, tablets, capsules, syrups and the like are preferable and can be safely administered orally.
  • the lactic acid strain of the present invention it is preferable to prepare a preparation using the dried microbial cells (live bacteria) of the lactic acid strain of the present invention.
  • a dry cell (live cell) of the lactic acid strain of the present invention can be obtained by a conventional method.
  • the lactic acid strain of the present invention is purely cultured, collected by a method such as centrifugation, and then added with an appropriate stabilizer. Obtained by freeze-drying.
  • the above-mentioned various preparations can be produced according to conventional methods, and together with the lactic acid strain of the present invention as the main agent, excipients, binders, disintegrants, coating agents, lubricants, stabilizers, flavoring agents, solubilizing aids It can be produced using known pharmaceutical additives that are usually used in the pharmaceutical formulation technical field such as drugs, lubricants, suspensions, and diluents.
  • the dose of each of the above preparations varies depending on the type and extent of the target disease. For example, about 1 mg to 2,000 mg per day as dry cells of the lactic acid strain of the present invention, depending on the symptoms It can be administered once or divided into several times.
  • the lactic acid strain of the present invention can be used by adding to confectionery such as tablet confectionery, gum, candy, and other general foods such as kimchi, and can provide a lactic acid bacteria-containing food. it can.
  • confectionery such as tablet confectionery, gum, candy, and other general foods such as kimchi
  • lactic acid bacteria-containing food it can.
  • the lactic acid strain of the present invention can be ingested in the form of a fermented food such as yogurt.
  • Such fermented foods include, for example, starter fungi for the production of York Noret, such as milk and sheep milk, Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus helveticus, Streptococcus thermophilus, It can be produced by inoculating the lactic acid strain of the present invention together with dairy lactic acid bacteria such as the above, and culturing mixed culture or mixing each of them after fermentation alone in a fermentation raw material.
  • starter fungi for the production of York Noret such as milk and sheep milk, Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus helveticus, Streptococcus thermophilus
  • the present inventor has found an additive or synergistic substance together with the lactic acid bacterial strain of the present invention.
  • the effect of the composition containing alcohols was investigated. As a result, it was found that when the sugar alcohol erythritol was used together with the lactic acid strain of the present invention, it exhibited a synergistic effect on the individual anti-cariogenic effect, respectively, in suppressing the production of insoluble glucan. (See Test Example 4).
  • Sugar alcohols such as xylitol, maltitol, sorbitol, and erythritol
  • oligosaccharides such as fructooligosaccharide, xylooligosaccharide, kestose, and raffinose
  • the synergistic effect or additive effect of sugar alcohols and oligosaccharides with lactic acid strains differs greatly depending on the selection of lactic acid strains.
  • sugar alcohols and oligosaccharides can be used alone or as a mixture in the live bacterial product or lactic acid bacteria-containing food of the present invention. These sugar alcohols and oligosaccharides can also serve as excipients or sweeteners. There are no particular restrictions on the types and proportions of these sugar alcohols and oligosaccharides, and any sugar alcohol or oligosaccharide can be used in any ratio. For example, in the case of tablet confectionery, any sugar alcohol or oligosaccharide can be selected by appropriately selecting conditions such as tablet composition, tableting pressure, tablet confectionery hardness, etc., using a conventional tableting technique. It is possible to easily manufacture products that contain
  • Test Example 1 LactobaciUus strain of Lactobacillus suitable for suppression of pathogenic bacteria in the oral cavity
  • MRS broth (Difico) medium was inoculated with 1 ⁇ 10 7 CFU / ml of 30 lactic acid bacteria belonging to the genus cto aciZ / ws shown in Table 1, and cultured at 37 ° C for 6 hours under aerobic conditions. The number of viable bacteria was measured by an agar plate method.
  • 1 ml of the culture solution of each strain at 6 hours above culture was anaerobic diluent (KH 2 PO 4 4.5 g / l, Na 2 HP0 4 6.0 g / l, cysteine hydrochloride 0.5 g / l, agar 0.5 g / l l, Tween 80 0.5 g / l), spread 0.1 ml of the diluted solution on MRS agar medium (agar plate with 1.5% agar added to MRS broth), 48 hours at 37 ° C After culturing under anaerobic conditions, The number of ronnies was measured and multiplied by the dilution factor to determine the number of viable bacteria. The results are shown in Table 1.
  • Table 1 shows ⁇ ⁇ zi ⁇ Vafe JCM 8525 strain (bacterial count: 1 X 10 6 CFU / ml) alone (control) or 10 ml of liquid medium supplemented with 0.7% glucose in GAM bouillon (Nissui) Each of the 30 strains of Lactobacillus was mixed and inoculated with each bacterium (number of bacteria: IX 10 6 CFU / ml). After culturing at 37 ° C for 12 hours under anaerobic culture conditions, the number of viable Porphyromonas gingivalis in the culture medium was measured by the agar plate method.
  • Porphyromonas gingivalis' ⁇ ⁇ ⁇ 3 ⁇ 4
  • the number of viable bacteria of Porphyromonas gingivalis during mixed culture with ⁇ cto ⁇ czT / s genus against the number (control, 100%) was determined.
  • EG-GM medium supplemented with gentamicin 10 gZml in EG agar medium (Nissui) was used as the selective medium, and only the number of Porphyromonas gingivals was grown. . That is,
  • ⁇ ⁇ z'ra is singly or mixed with each of the Lactobacillus strains (30 strains) above, each lml was collected and serially diluted with the anaerobic diluent, Spread 0.1 ml on the above EG-GM agar plate with a large rod, and incubate for 48-72 hours at 37 ° C. Count the number of colonies obtained, and multiply the dilution factor to determine the number of viable bacteria. .
  • the survival rate (%) is shown in Table 1. Therefore, the inhibition rate (%) of actoba T / iw against? ⁇ afo is expressed by the value (° / o) obtained by subtracting the survival rate (%) of P. gingivalis from Control (100%). Will be. H gingivalis cocoons in mixed cultures
  • 3 ⁇ 4re tococcws nwfei2s counts by agar plate method include TCYSB agar (tripty case soger (BBL) 40 g / l, cystine 0.3 g / l, yeast extract 5 g / l, sucrose 200 g / l, By using agar (5 g / l, bacitracin 10 U / ml) as a selective medium, Streptococcus mutans was contributed and the number of bacteria was measured.
  • TCYSB agar tripty case soger (BBL) 40 g / l, cystine 0.3 g / l, yeast extract 5 g / l, sucrose 200 g / l
  • BBL tripty case soger
  • Streptococcus mutans viable count (control, 100%) vs. Streptococcus / Hwaas viable count (%) in Streptococcus mutans viable count (control, 100%) )
  • the survival rate (%) is shown in Table 1. Therefore, the inhibition rate (%) of Lactobacillus against S. iz a ⁇ is from Control (100%). It is expressed by the value (%) obtained by subtracting the survival rate (%).
  • Table 1 shows the ratio of the amount of insoluble glucan measured when mixed with each Lactobacillus genus to the amount of insoluble glucan measured when Streptococcus mutans was cultured alone (insoluble glucan production rate,%). Shown in Therefore, the inhibition rate (%) of L & ctobacillus ⁇ ⁇ against S. mutans is the value obtained by subtracting the production rate (%) of insoluble glucan produced by S. mutans from Control (100%) (%) It will be represented by
  • Control is the number of bacteria obtained by culturing each of JR gingivalis and S. mutans separately, and is shown as 100 (%). From the results shown in Table 1, among the Lactobacil ⁇ us genus strains, the most proliferative, and periodontal It was revealed that (26) strain Lactobacillus salivariusTl 2711 was the most excellent strain that suppressed the growth of Streptococcus mutans, a fasting fungus, and produced strong insoluble glucan. Test Example 2: In vivo mouse mouth of Lactobacillus salivarius TI 2711 strain Porphyromonas gingivalis (causing periodontal disease) and Streptococcus mutans
  • 1 x 10 9 CFU / 0.5ml of Porphyromonas gingivalis JCM8525, a periodontal disease-causing bacterium was inoculated into the oral cavity 3 times in total for 3 consecutive days in the post-infection administration group and the Control group, and one week after the infection was established
  • the Lactobacillus salivarius TI 2711 strain (the number of bacteria 1 ⁇ 10 9 CFU / 0.5 ml), which is the lactic acid strain of the present invention, was administered to the post-infection administration group for 3 consecutive days, 3 times in total.
  • 1 x 10 9 CFU / 0.5ml of Streptococcus mutans MT8148, a caries-causing bacterium was inoculated into the oral cavity three times in total for 3 consecutive days in the post-infection administration group and the Control group, and further, 1 week after Lactobacillus salivai'ius mil (bacterial count 1 ⁇ 10 9 CFU / 0.5 ml), which is the lactic acid strain of the present invention, was administered to the post-administration group for 3 consecutive days for a total of 3 times.
  • the number of bacteria of SetoiWi i / s / Hi ⁇ aas was measured in the same manner as in item (3) of Test Example 1.
  • Lactic acid strain of the present invention Prepare confections containing 140 mg of salivariusTl 2711 lyophilized bacterial powder per tablet (bacteria count 1 X 10 8 CFU / g), 5 tablets at a time, 5 times a day for a total of 25 tablets between meals I took it. They were taken every day for a period of 2 months, and saliva was collected, halitosis was measured, and doctors were interviewed.
  • FIG 11 shows an overview of the clinical trial protocol for this study.
  • Lactobacilli Modified LBS agar medium
  • BPAR anaerobic gram-negative black koji mold
  • Figure 3 shows the total bacterial count results obtained. As shown in Fig. 3, there was no significant difference in the total number of bacteria in the human oral cavity before taking tablet candy and after taking it for 4 weeks.
  • Fig. 4 shows the results of the number of oral periodontal disease bacteria obtained. As shown in Figure 4, before taking There are 8 people with the number of bacteria below the detection limit, and the average value was 10 6 6 ⁇ 1 ' 3 CFU / total saliva, but 4 weeks after taking Those with numbers increased significantly to 30, and the average value was 10 5 ' 3 ⁇ 1 ⁇ 6 CFU / total saliva (PO.0001). Therefore, it was shown that periodontal disease bacteria, the pathogen, were sterilized and suppressed by taking Lactobacillus salivai'iusTl 2711.
  • Fig. 5 shows the results of the number of caries bacteria obtained. As shown in Fig. 5, there was no significant difference in the number of bacteria in the saliva of Streptococcus mutans before and after taking.
  • Fig. 7 shows the measurement results of saliva pH fluctuations.
  • the pH fluctuation pH fluctuation range
  • the pH variation became small and the average value was ⁇ 7.3 ⁇ 0.2, which is almost the same as the pH of blood. That is, the pH of saliva is maintained normally. This phenomenon was observed after 8 weeks, the pH was maintained normally, and there was very little variation.
  • Fig. 8 shows the results of measurement of the amount of insoluble glucan produced in saliva.
  • the average amount of insoluble glucan in saliva before monthly saliva was 9.9 ⁇ 6.0 mg / total saliva, but the average value was 7.6 ⁇ 5.8 at 4 weeks after taking the drug. mg / total saliva (PO.05), a statistically significant difference.
  • the mean value was 4.2 ⁇ 2.2 mg / total saliva (P 0.001), and Dose Response Correlation was observed after taking Lactobacillus salivarius TI 2711. This is part of the evidence that constitutes the scientific basis for the effectiveness of the lactic acid strains of the invention demonstrated by this clinical trial.
  • the lactic acid strain of the present invention is insoluble, which is a major causative substance of dental plaque (Dental plaque).
  • Dental plaque can stop the production of sex glucan, cut off the habitat of anaerobic periodontal and carious bacteria from the beginning, and can be a means to suppress the progression, recurrence and relapse of chronic infections such as caries and periodontal disease It has been found.
  • Lactobacillus salivarius TI 2711 is taken over a long period of time, as mentioned above, salivary ⁇ ⁇ becomes normal and it is difficult to form strong plaque. It was strongly suggested that this is useful for preventing caries and periodontal disease and treating mild cases.
  • halitosis protein degradation by periodontal disease bacteria.
  • periodontal disease bacteria have extremely high activity of proteolytic enzymes (proteases), so they easily break down proteins such as oral food waste, which is one of the nutrient sources of the bacteria, and 3 ⁇ 4S, CH 3 SH and others It generates odor-causing substances that are volatile sulfur compounds such as Thiols and Sulfides.
  • the RU value of the bad breath of these 20 people was 164 ⁇ 96 ppb (average soil standard error) before taking, but it decreased to 94 ⁇ 21 ppb 4 weeks after taking A very significant difference (P ⁇ 0.005) was observed.
  • erythritol was the substance that inhibited the production of the most insoluble glucan among sugar alcohols and oligosaccharides, and erythritol alone inhibited about 60%, but in addition, Lactobacillus sali temporary ius l 2711 strain As a result of inoculating and incubating, the additive or synergistic effect was observed, showing an inhibition rate of about 90% when Lactobacillus salivarius non-inoculated insoluble 'I' raw glucan was taken as 100%. The result was obtained. Table 2
  • Example 1 Preparation of dry cells of Lactobacillus salivarius ⁇ 2711 strain
  • the Lactobacillus salivarius TI 2711 strain was inoculated into a brick liver liquid medium containing 0.3% calcium carbonate, and then statically cultured at 37 ° C for 18 hours. After completion of the culture, centrifugation was performed at 7,000 rpm for 15 minutes to obtain concentrated bacterial cells with 1/100 volume of the culture solution.
  • the concentrated cells are mixed with a dispersion medium containing 5% (by weight) sodium glutamate, 5% (by weight) soluble starch, and 5% (by weight) sucrose, and adjusted to pH 7.0%. Freeze-drying was performed after freezing at C or lower. The resulting lyophilized powder was powdered on a 60 mesh sieve to prepare the lactic acid powder of the present invention. Regarding the storage stability of this bacterium, no decrease in the number of bacteria was observed even when the lactic acid bacteria powder was stored at room temperature 24 ° (:, 10 months, sealed condition (aluminum laminate bag). Manufacture of live pharmaceutical preparations (tablets)
  • Example 3 Production of food containing lactic acid bacteria (tablet confectionery)
  • 95g of uncoated tablets of 300mg per tablet were produced by compression molding with a tablet machine.
  • Example 4 Production of food (gum) containing lactic acid bacteria
  • Lactobacillus salivarius TI 2711 strain dry powder 10g (bacteria count, 5 X 10 9 CFU / equivalent) Erythritol 160g, sorbitol 160g, peppermint oil 20g as a fragrance and gum base 150g
  • erythritol and sorbitol which are sweeteners, and acto> aiy: Z / ws
  • a mixture of 2711 dry bacteria powder was mixed evenly when kneaded with gum base and mixed uniformly. Finally, peppermint oil was added to add flavor and kneaded uniformly.
  • Example 5 Production of food containing lactic acid bacteria (fermented milk)
  • Lactobacillus acidophilus which is a starter fungus of fermented milk, was inoculated into a reduced defatted medium containing 23 g of skim milk powder, 1.0 g of yeast extract and 0.06 g of ascorbic acid, and statically cultured at 37 ° C for 16 hours was used as a bulk starter.
  • oral flora can be normalized by taking or ingesting the strain of the present invention, and as a result, periodontal bacteria and caries can be suppressed, and thus the occurrence of halitosis can be suppressed.

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  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Inorganic Chemistry (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

L'invention concerne des préparations cellulaires vitales ou des aliments contenant une bactérie lactique, à savoir le Lactobacillus salivarius, comme principe actif. Ces préparations et aliments permettent d'éviter l'occurrence, la récurrence ou la recrudescence de parodontopathies et/ou caries provoquées par des bactéries associées aux parodontopathies et/ou caries et de régulariser la flore orale, de manière à éviter les problèmes de mauvaise haleine et à maintenir la salive dans une gamme de pH physiologiquement normale.
PCT/JP2002/003177 2002-03-29 2002-03-29 Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif WO2003082027A1 (fr)

Priority Applications (10)

Application Number Priority Date Filing Date Title
PCT/JP2002/003177 WO2003082027A1 (fr) 2002-03-29 2002-03-29 Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif
CNB02828691XA CN1309316C (zh) 2002-03-29 2002-03-29 含乳酸杆菌作为活性成分的活细菌制剂以及含有乳酸杆菌的食品
DK02708726.1T DK1498039T3 (da) 2002-03-29 2002-03-29 Vitale cellepræparater, som indeholder mælkesyrebakterie som aktivt stof, og mælkesyrebakterieholdige fødevareprodukter
US10/508,340 US7674613B2 (en) 2002-03-29 2002-03-29 Live bacterium preparation containing lactic acid bacterium as active ingredient and food containing lactic acid bacterium
KR1020047015110A KR100843020B1 (ko) 2002-03-29 2002-03-29 유산균을 유효성분으로 하는 생균제제 및 유산균 함유 식품
EP02708726A EP1498039B1 (fr) 2002-03-29 2002-03-29 Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif
JP2003579579A JP4203855B2 (ja) 2002-03-29 2002-03-29 乳酸菌を有効成分とする生菌製剤および乳酸菌含有食品
AU2002243005A AU2002243005A1 (en) 2002-03-29 2002-03-29 Vital cell preparations containing lactic acid bacterium as the active ingredient and lactic acid bacterium-containing foods
TW92107146A TW200305370A (en) 2002-03-29 2003-03-28 Vital cell preparations containing lactic acid bacterium as the active ingredient and lactic acid bacterium-containing foods
HK05106059A HK1073230A1 (en) 2002-03-29 2005-07-18 Live bacterium preparation containing lactic acid bacterium as active ingredient and food containinglactic acid bacterium

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2002/003177 WO2003082027A1 (fr) 2002-03-29 2002-03-29 Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif

Publications (1)

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WO2003082027A1 true WO2003082027A1 (fr) 2003-10-09

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PCT/JP2002/003177 WO2003082027A1 (fr) 2002-03-29 2002-03-29 Preparations cellulaires vitales et aliments contenant une bacterie lactique comme principe actif

Country Status (10)

Country Link
US (1) US7674613B2 (fr)
EP (1) EP1498039B1 (fr)
JP (1) JP4203855B2 (fr)
KR (1) KR100843020B1 (fr)
CN (1) CN1309316C (fr)
AU (1) AU2002243005A1 (fr)
DK (1) DK1498039T3 (fr)
HK (1) HK1073230A1 (fr)
TW (1) TW200305370A (fr)
WO (1) WO2003082027A1 (fr)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006022471A1 (fr) * 2004-08-25 2006-03-02 Jong Suk Oh Bacterie d'acide lactique arretant la mauvaise haleine
WO2006033949A1 (fr) * 2004-09-21 2006-03-30 The Procter & Gamble Company Lactobacillus iners destines a lutter contre les mauvaises odeurs humaines
JP2008502714A (ja) * 2004-06-14 2008-01-31 バイオガイア・エイビー 歯肉出血を減少させ、口内炎症を軽減するための乳酸菌の使用
JP2010124772A (ja) * 2008-11-28 2010-06-10 Lion Corp 乳酸菌及びこれを含有する口腔用組成物、ならびに口腔内バイオフィルム形成抑制剤、ポルフィロモナス・ジンジバリス及び／又はフゾバクテリウム・ヌクレアタムの生育阻害剤、う蝕予防剤、歯周病予防・治療剤及び口臭改善・予防剤
WO2011007584A1 (fr) 2009-07-16 2011-01-20 国立大学法人広島大学 Agent prophylactique, d'amélioration et thérapeutique pour maladies buccales
JP2012092094A (ja) * 2010-09-30 2012-05-17 Wakamoto Pharmaceutical Co Ltd 油脂中に乳酸菌を含有する口腔用組成物
JP2012532593A (ja) * 2009-07-10 2012-12-20 バイオガイア・エイ・ビー 経口補水液用粉末と混合した凍結乾燥乳酸菌を保存するための製品
WO2014135300A1 (fr) 2013-03-04 2014-09-12 Swiss Stem Cell Foundation Système d'extraction de cellules dans un échantillon de tissu
JP2017088580A (ja) * 2015-11-16 2017-05-25 欽也 斉藤 口内虫歯予防粉及び口内虫歯予防粉の製造方法
WO2018207924A1 (fr) * 2017-05-12 2018-11-15 キッコーマン株式会社 Procédé de production d'une bactérie lactique riche en arn double brin et ladite bactérie lactique
JP6487106B1 (ja) * 2018-10-15 2019-03-20 株式会社湖池屋 免疫機能向上用食品組成物
JP2020080785A (ja) * 2018-11-29 2020-06-04 雪印メグミルク株式会社 歯周病予防用組成物

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2133414A1 (fr) 2008-06-11 2009-12-16 Basf Se Utilisations et procédés pour prévenir et/ou traiter la mauvaise haleine
CN102311932B (zh) * 2010-07-07 2014-03-19 生展生物科技股份有限公司 改善口腔内细菌群的唾液乳酸杆菌及其保健组合物
CN102618452B (zh) * 2011-02-01 2014-06-25 任发政 唾液乳杆菌及其代谢物的制备方法和组合物以及应用
WO2013021957A1 (fr) 2011-08-05 2013-02-14 株式会社ヤクルト本社 Agent prophylactique ou thérapeutique pour des maladies buccales
CN104825380A (zh) * 2015-04-21 2015-08-12 溧水区人民医院 一种快速恢复口腔正常菌群的护理液及其制备方法
KR102095339B1 (ko) * 2017-06-01 2020-05-27 한국생명공학연구원 신규한 락토바실러스 류테리 균주 및 이를 유효성분으로 포함하는 치주질환 예방 또는 치료용 조성물
CA3079101A1 (fr) * 2017-11-16 2019-05-23 The Procter & Gamble Company Dispositif de demonstration de produit et procede associe
US11147845B2 (en) 2018-07-16 2021-10-19 ProBiora Health, LLC Foods containing beneficial oral bacteria
KR102001074B1 (ko) * 2018-12-07 2019-07-18 주식회사 메디오젠 충치 억제 활성을 갖는 유산균 조성물
KR102230517B1 (ko) * 2019-07-09 2021-03-22 주식회사 메디오젠 충치 억제 활성을 갖는 락토바실러스 살리바리우스를 포함하는 조성물
KR102210092B1 (ko) * 2019-07-09 2021-02-01 주식회사 메디오젠 충치 억제 활성을 갖는 락토바실러스 루테리 mg505 를 포함하는 조성물
KR102649531B1 (ko) * 2019-12-26 2024-03-22 여장욱 동물용 프로바이오틱스 페이스트 및 그 제조 방법
CN112126605A (zh) * 2020-10-12 2020-12-25 镇江市天益生物科技有限公司 一种唾液乳杆菌及其应用
CN114053313B (zh) * 2022-01-17 2022-04-01 中科嘉亿营养医学(山东)微生态研究院有限公司 一种唾液乳杆菌jyls-372在制备解酒护肝产品中的应用
CN115040465B (zh) * 2022-07-06 2023-01-24 浙江爱尚日用品有限公司 一种有效祛除牙斑、牙垢的增白牙膏

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0154549A2 (fr) * 1984-03-09 1985-09-11 Kabushiki Kaisya Advance Agent anticarieux ou antipériodontique
JPH107577A (ja) * 1996-06-17 1998-01-13 Yakult Honsha Co Ltd 血糖降下剤
WO2001010233A1 (fr) * 1999-08-03 2001-02-15 Kabushiki Kaisha Yakult Honsha Boissons et aliments a base de lait fermente et leur procede de production

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6191126A (ja) 1984-10-09 1986-05-09 Advance Res & Dev Co Ltd 抗歯周症剤
DE3439094A1 (de) * 1984-10-25 1986-05-07 Henkel KGaA, 4000 Düsseldorf Mund- und zahnpflegemittel
KR100266752B1 (ko) * 1997-08-07 2001-03-02 오종석 인체 구강내 치태형성을 억제하는 신규한 유산균
JP3648115B2 (ja) 2000-01-14 2005-05-18 株式会社ヤクルト本社 乳酸菌の培養方法及び飲食品
DE60137235D1 (de) * 2000-08-25 2009-02-12 Wakamoto Pharma Co Ltd L. salivarius-Stamm enthaltende probiotische Produkte

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0154549A2 (fr) * 1984-03-09 1985-09-11 Kabushiki Kaisya Advance Agent anticarieux ou antipériodontique
JPH107577A (ja) * 1996-06-17 1998-01-13 Yakult Honsha Co Ltd 血糖降下剤
WO2001010233A1 (fr) * 1999-08-03 2001-02-15 Kabushiki Kaisha Yakult Honsha Boissons et aliments a base de lait fermente et leur procede de production

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP1498039A4 *

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008502714A (ja) * 2004-06-14 2008-01-31 バイオガイア・エイビー 歯肉出血を減少させ、口内炎症を軽減するための乳酸菌の使用
CN102851350A (zh) * 2004-06-14 2013-01-02 生命大地女神有限公司 使用乳酸杆菌减少牙龈出血和减轻口腔炎症
CN102851350B (zh) * 2004-06-14 2015-12-16 生命大地女神有限公司 使用乳酸杆菌减少牙龈出血和减轻口腔炎症
WO2006022471A1 (fr) * 2004-08-25 2006-03-02 Jong Suk Oh Bacterie d'acide lactique arretant la mauvaise haleine
WO2006033949A1 (fr) * 2004-09-21 2006-03-30 The Procter & Gamble Company Lactobacillus iners destines a lutter contre les mauvaises odeurs humaines
JP2010124772A (ja) * 2008-11-28 2010-06-10 Lion Corp 乳酸菌及びこれを含有する口腔用組成物、ならびに口腔内バイオフィルム形成抑制剤、ポルフィロモナス・ジンジバリス及び／又はフゾバクテリウム・ヌクレアタムの生育阻害剤、う蝕予防剤、歯周病予防・治療剤及び口臭改善・予防剤
JP2012532593A (ja) * 2009-07-10 2012-12-20 バイオガイア・エイ・ビー 経口補水液用粉末と混合した凍結乾燥乳酸菌を保存するための製品
US10143712B2 (en) 2009-07-16 2018-12-04 Hiroshima University Prophylactic, ameliorating or therapeutic agent for oral diseases
WO2011007584A1 (fr) 2009-07-16 2011-01-20 国立大学法人広島大学 Agent prophylactique, d'amélioration et thérapeutique pour maladies buccales
KR20120035923A (ko) 2009-07-16 2012-04-16 고쿠리츠다이가쿠호진 히로시마다이가쿠 구강내 질환의 예방, 개선 또는 치료제
CN102470151A (zh) * 2009-07-16 2012-05-23 国立大学法人广岛大学 口腔内疾病的预防、改善或治疗剂
JP2012092094A (ja) * 2010-09-30 2012-05-17 Wakamoto Pharmaceutical Co Ltd 油脂中に乳酸菌を含有する口腔用組成物
WO2014135300A1 (fr) 2013-03-04 2014-09-12 Swiss Stem Cell Foundation Système d'extraction de cellules dans un échantillon de tissu
JP2017088580A (ja) * 2015-11-16 2017-05-25 欽也 斉藤 口内虫歯予防粉及び口内虫歯予防粉の製造方法
WO2018207924A1 (fr) * 2017-05-12 2018-11-15 キッコーマン株式会社 Procédé de production d'une bactérie lactique riche en arn double brin et ladite bactérie lactique
JP6480090B1 (ja) * 2017-05-12 2019-03-06 キッコーマン株式会社 二本鎖ｒｎａ高含有乳酸菌の製造方法及び該乳酸菌
US11518976B2 (en) 2017-05-12 2022-12-06 Kikkoman Corporation Method for producing double-stranded RNA-rich lactic acid bacterium, and said lactic acid bacterium
JP6487106B1 (ja) * 2018-10-15 2019-03-20 株式会社湖池屋 免疫機能向上用食品組成物
JP2020061945A (ja) * 2018-10-15 2020-04-23 株式会社湖池屋 免疫機能向上用食品組成物
JP2020080785A (ja) * 2018-11-29 2020-06-04 雪印メグミルク株式会社 歯周病予防用組成物
JP7160652B2 (ja) 2018-11-29 2022-10-25 雪印メグミルク株式会社 歯周病予防用組成物

Also Published As

Publication number Publication date
CN1625345A (zh) 2005-06-08
CN1309316C (zh) 2007-04-11
JP4203855B2 (ja) 2009-01-07
HK1073230A1 (en) 2005-09-30
JPWO2003082027A1 (ja) 2005-07-28
TWI342750B (fr) 2011-06-01
KR20040111434A (ko) 2004-12-31
TW200305370A (en) 2003-11-01
EP1498039A4 (fr) 2006-03-29
DK1498039T3 (da) 2012-10-15
US20070071737A1 (en) 2007-03-29
EP1498039B1 (fr) 2012-10-03
KR100843020B1 (ko) 2008-07-01
EP1498039A8 (fr) 2005-06-22
US7674613B2 (en) 2010-03-09
AU2002243005A1 (en) 2003-10-13
EP1498039A1 (fr) 2005-01-19

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